Liquid chromatography–electrospray mass spectrometry and tandem mass spectrometry of ceramides

• Published in: Analytica chimica acta Vol. 403; no. 1; pp. 295 - 303
• Main Authors: Raith, Klaus, Neubert, Reinhard H.H
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 03.01.2000; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Ceramides; Electrospray; Fundamental and applied biological sciences. Psychology; LC/MS; Lipids; MS/MS; Other biological molecules; Sphingolipids; LC/MS; Ceramides; MS/MS; Electrospray; Ion trap; Collisional activation; Biochemical analysis; Multicomponent analysis; Coupled method; Mass spectrometry MS/MS; Ceramide; Sphingolipid; Liquid chromatography; Quadrupole spectrometer; Quantitative analysis
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/S0003-2670(99)00661-3

A quantitative liquid chromatography–mass spectrometry (LC/MS) method designed for the rapid, sensitive and specific analytical determination of all kinds of ceramides is described. LC separation was carried out on a RP 18 column (2 mm i.d., 3 μm particle size) using an isocratic mobile phase (methanol/tetrahydrofuran 97 : 3) in less than 8 min. LC/MS and LC/MS/MS were performed on an ion trap instrument and were compared to results obtained using a single quadrupole instrument. In the negative ionization mode a limit of detection of approx. 500 pg/ml was reached. The quantification using external standards showed good linearity over three orders of magnitude and a relative standard deviation of 3%. The use of Tandem MS and MS 3 for obtaining structural information was demonstrated on the ion trap instrument and compared to results achieved on a triple quadrupole instrument.